Modified isocratic capillary electrophoresis detection of cell-free DNA in semen.

ABSTRACT

PURPOSE: The objectives were i) to analyze semen for the presence of cell-free DNA and ii) to determine the association between sperm parameters and cell-free DNA. METHODS: Cell-free DNA in semen (N = 25 cases) were detected using the modified capillary gel electrophoresis (CE) procedure. SYBR-Gold was used to stain high (12 Kb) and low (1 Kb) molecular weight DNA fragments and the images analyzed. RESULTS: The quantity of low-molecular weight cell-free DNA was positively correlated to rapid progression, curvilinear velocity (>40 microm/s), normal strict morphology and capacitation index. High-molecular weight cell-free DNA intensity index was negatively correlated to post-wash hyperactivation. Sperm concentration was not related to cell-free DNA quantity. The sperm freezing process did not increase cell-free DNA but reduced the more labile low-molecular weight cell-free DNA. CONCLUSIONS: Cell-free DNA present in semen was correlated to important sperm parameters linked to normal sperm function. The data suggested the possible use of cell-free DNA as a marker of semen quality. This study reports on the novel finding of cell-free DNA released along with sperm during each ejaculation.